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Preservation of Bovine Spermatozoa at –79 and –196° C.¹

Department of Animal Industries, University of Connecticut, Storrs

ABSTRACT

Studies were conducted primarily to determine the effect of storage temperatures of –79 and –196° C. on fertility of bovine spermatozoa.

A total of 145 ejaculates from 43 bulls of the Holstein, Jersey, Guernsey, Angus, Ayrshire, and Brown Swiss breeds were frozen in an egg yolk-citrate extender. The 60- to 90-day per cent nonreturn (NR) rate from 3,214 first-service inseminations with liquid nitrogen (LN) stored semen was 70.6, compared to 3,091 first services with a NR rate of 67.8% from DI storage. The weighted mean difference in per cent NR of 2.9 was statistically significant (P < 0.02).

Breeding efficiencies were obtained on second-service inseminations from the two treatments. The number of services and 60- to 90-day per cent NR was 1,132, 68.1 versus 1,143, 66.9 for LN and dry ice-alcohol (DI) stored semen, respectively. The weighted mean difference of 1.1% NR in favor of LN was not significant (P = 0.05).

In a second field trial, semen from nine Holstein bulls was frozen in LN, to –80° C, and then divided equally for storage at –79 and –196° C. A total of 299 first-service cows were bred with semen stored in LN for a 60- to 90-day NR of 73.2%, as compared to a NR rate of 67.6% on 330 first services to DI stored semen. There was a weighted mean difference of 5.3% in favor of LN. The difference was not statistically significant at P = 0.05.

Combined data from this station included a total of 9,742 first-service cows, 4,951 with semen stored in LN versus 4,791 with semen stored in DI. The 60- to 90-day per cent NR was 71.1, compared to 68.4 for LN and DI, respectively. There was a significant difference (P < 0.01), indicating that LN, under the conditions of these studies, is a superior refrigerant for storage of bovine spermatozoa.

Thirteen measurements were collected on each of 132 ejaculates. These characteristics were correlated with fertility of semen stored in DI and LN. All correlations were too low to be useful in predicting the fertility of frozen semen.

¹ Supported in part by grants-in-aid from Chas. H. Hood Dairy Foundation, Connecticut Artificial Breeding Association, National Association of Artificial Breeders, New England Artificial Breeding Council, New England Selective Breeding Association, and the New Hampshire-Vermont Breeding Association. Portions of these data were presented at the Eastern Division of the A.D.S.A. Annual Meeting, August 7–8, 1961, University of Massachusetts, Amherst.